Figure 5
Figure 5. Longer blockade of anergic signaling induces apoptosis in the pERK(+) subset. (A) Leukemic cells were left untreated or treated for 48 hours with 10μM U0126, and cell viability was measured with the use of a luminescent-based cell viability assay. Each value from treated samples was normalized to the untreated control, and samples were grouped on the basis of the ERK1/2 activation status. Data were analyzed by using Mann-Whitney U test (P value is indicated). (B) Leukemic cells were left untreated or treated for 48 hours with 10μM CI1040, and cell viability was measured with a luminescent-based cell viability assay. Each value from treated samples was normalized to the untreated control, and samples were grouped on the basis of the ERK1/2 activation status. Data were analyzed by using Mann-Whitney U test (P value is indicated). (C) Leukemic cells were left untreated or treated for 48 hours with 10μM NMS6E and cell viability was measured by the use of a luminescent-based cell viability assay. Each value from treated samples was normalized to the untreated control, and samples were grouped on the basis of ERK1/2 activation status. Data were analyzed by using Mann-Whitney U test (P value is indicated). (D-E) Leukemic cells were left untreated or treated for 48 hours with 10μM 11R-VIVIT, and cell viability was measured by the use of luminescent-based cell viability assay. Each value from treated samples was normalized to the untreated control, and samples were grouped on the basis of ERK1/2 (D) or NF-ATc1 (E) activation status. Data were analyzed with the Mann-Whitney U test (P value is indicated). (F) ERK1/2 and NF-ATc1 inhibitors do not exert any synergistic effect. Samples from 9 anergic CLL patients were treated with 10μM each inhibitor (U0126, NMS6E, VIVIT) or with a MAPK inhibitor in combination with VIVIT for 48 hours, and cell viability was measured. Viability is expressed as percentage of the untreated control.

Longer blockade of anergic signaling induces apoptosis in the pERK(+) subset. (A) Leukemic cells were left untreated or treated for 48 hours with 10μM U0126, and cell viability was measured with the use of a luminescent-based cell viability assay. Each value from treated samples was normalized to the untreated control, and samples were grouped on the basis of the ERK1/2 activation status. Data were analyzed by using Mann-Whitney U test (P value is indicated). (B) Leukemic cells were left untreated or treated for 48 hours with 10μM CI1040, and cell viability was measured with a luminescent-based cell viability assay. Each value from treated samples was normalized to the untreated control, and samples were grouped on the basis of the ERK1/2 activation status. Data were analyzed by using Mann-Whitney U test (P value is indicated). (C) Leukemic cells were left untreated or treated for 48 hours with 10μM NMS6E and cell viability was measured by the use of a luminescent-based cell viability assay. Each value from treated samples was normalized to the untreated control, and samples were grouped on the basis of ERK1/2 activation status. Data were analyzed by using Mann-Whitney U test (P value is indicated). (D-E) Leukemic cells were left untreated or treated for 48 hours with 10μM 11R-VIVIT, and cell viability was measured by the use of luminescent-based cell viability assay. Each value from treated samples was normalized to the untreated control, and samples were grouped on the basis of ERK1/2 (D) or NF-ATc1 (E) activation status. Data were analyzed with the Mann-Whitney U test (P value is indicated). (F) ERK1/2 and NF-ATc1 inhibitors do not exert any synergistic effect. Samples from 9 anergic CLL patients were treated with 10μM each inhibitor (U0126, NMS6E, VIVIT) or with a MAPK inhibitor in combination with VIVIT for 48 hours, and cell viability was measured. Viability is expressed as percentage of the untreated control.

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